LOCUS       sequence-328394-        8203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression vector for FLAG-tagged DPP8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8203)
  AUTHORS   Hollingsworth LR, Sharif H, Griswold AR, Fontana P, Mintseris J, 
            Dagbay KB, Paulo JA, Gygi SP, Bachovchin DA, Wu H
  TITLE     DPP9 sequesters the C terminus of NLRP1 to repress inflammasome 
            activation.
  JOURNAL   Nature. 2021 Mar 17. pii: 10.1038/s41586-021-03350-4. doi: 
            10.1038/s41586-021-03350-4.
  PUBMED    33731932
REFERENCE   2  (bases 1 to 8203)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2021 Mar 17. pii: 10.1038/s41586-021-03350-4. doi: 
            10.1038/s41586-021-03350-4."
COMMENT     Sequence Label: pcDNA3.1 LIC 6A FLAG-TEV-DPP8
FEATURES             Location/Qualifiers
     source          1..8203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      929..938
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             944..967
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             989..1009
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     promoter        complement(3756..3774)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3757..3774)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3794..3811)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3800..4024
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4070..4498
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4157..4176)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4367..4388
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4507..4527)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4512..4841
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4692..4827
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4754..4773
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4908..5702
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4962..4981)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5572..5591
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5876..5997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5913..5932)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5967..5986
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6046..6062)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6046..6062)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6059..6081)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6070..6086
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6094..6100,6101..6118,6119..6124))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6139..6160
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6277..6294)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6448..7036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6528..6547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7207..7998,7999..8067))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7830..7849
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8068..8172)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccctacttcc aatccaatgc caccatggat atcgactaca aggacgatga
      961 cgacaaggat tacgatatcc caacgaccga aaacctgtat tttcagggcg ccatgggatc
     1021 catgtggaag cgttccgagc agatgaagat caagtccggc aagtgcaaca tggctgctgc
     1081 tatggagact gagcagctgg gtgtcgagat cttcgagact gctgactgcg aggagaacat
     1141 cgagtcccag gaccgtccca agctggagcc cttctacgtg gagcgttact cctggtccca
     1201 gctgaagaag ctgctggctg acacccgcaa gtaccacggt tacatgatgg ctaaggctcc
     1261 ccacgacttc atgttcgtga agcgtaacga ccccgacggt ccccactccg accgtatcta
     1321 ctacctggct atgtccggcg agaaccgcga gaacaccctg ttctactccg agatccccaa
     1381 gaccatcaac cgtgctgctg tgctgatgct gtcctggaag cccctgctgg atctcttcca
     1441 agctaccctg gactacggca tgtactcccg cgaggaggag ctgctgcgcg agcgcaagcg
     1501 tatcggcacc gtgggtatcg ctagctacga ctaccaccag ggttccggca ccttcctgtt
     1561 ccaggccggt tccggtatct accacgtgaa ggacggtggt ccccagggtt tcactcagca
     1621 gcccctgcgt cccaacctgg tggagacttc ctgccccaac atccgtatgg accccaagct
     1681 gtgccccgct gaccccgact ggatcgcttt catccactcc aacgacatct ggatctccaa
     1741 catcgtgact cgcgaggagc gtcgtctgac ctacgtgcac aacgagctgg ctaacatgga
     1801 ggaggacgct cgttccgctg gtgtcgctac cttcgtgctg caggaggagt tcgaccgtta
     1861 ctccggttac tggtggtgcc ccaaggctga gactaccccc tccggtggca agatcctgcg
     1921 tatcctgtac gaggagaacg acgagagcga ggtggagatc atccacgtga cctcccccat
     1981 gctggagact cgtcgtgctg actccttccg ttaccccaag accggcaccg ctaaccccaa
     2041 ggtcaccttc aagatgtccg agatcatgat cgacgctgag ggtcgtatca tcgacgtgat
     2101 cgacaaggag ctgatccagc ccttcgagat cctgttcgag ggtgtcgagt acattgctcg
     2161 tgctggctgg acccccgagg gcaagtacgc ttggtctatt ctcctcgata gatctcagac
     2221 ccgtctgcag atcgtcctca tttctcctga actgttcatc cccgtggagg acgacgtgat
     2281 ggagcgtcag cgtctgatcg agtccgtgcc cgactccgtg acccccctga tcatctacga
     2341 ggagactact gacatctgga tcaacatcca cgacatcttc cacgtgttcc cccagtctca
     2401 cgaagaagaa attgagttca tcttcgcttc cgagtgcaag accggtttcc gtcacctgta
     2461 caagatcacc tccatcctga aggagtccaa gtacaagcgt tcctccggcg gtctgcccgc
     2521 tccctccgac ttcaagtgcc ccatcaagga ggagatcgct atcacctccg gcgagtggga
     2581 ggtgctgggt cgtcacggtt ccaacatcca ggtggacgag gtccgccgtc tggtgtactt
     2641 cgagggcacc aaggactccc ccctggagca ccacctgtac gtggtgtcct acgtgaaccc
     2701 cggcgaggtc acccgtctga ccgaccgtgg ttactcccac tcctgctgca tctcccagca
     2761 ctgcgacttt ttcatctcca agtactccaa ccagaagaac ccccactgcg tcagcctcta
     2821 caagctgtcc tcccccgagg acgaccccac ctgcaagacc aaggagttct gggctaccat
     2881 cctggactcc gctggtcctc tgcccgacta caccccccct gagatcttct ccttcgagtc
     2941 caccaccggt ttcaccctgt acggcatgct gtacaagccc cacgacctgc agcccggcaa
     3001 gaagtacccc accgtcctgt tcatctacgg tggccctcag gtgcagctgg tcaacaaccg
     3061 tttcaagggc gtgaagtact tccgtctgaa caccctggct tctctcggat acgtcgtggt
     3121 cgtcatcgac aaccgtggtt cttgccaccg tggtctgaag ttcgagggcg ccttcaagta
     3181 caagatgggc cagatcgaga tcgacgacca ggtggagggc ctgcagtacc tggcctcccg
     3241 ttacgacttc atcgacctgg accgtgtggg tatccacggc tggtcctacg gtggttacct
     3301 gtccctgatg gctctgatgc agcgttccga catcttccgt gtggctatcg ctggtgctcc
     3361 cgtgaccctg tggatcttct acgacaccgg ttacaccgag cgttacatgg gtcaccccga
     3421 ccagaacgag cagggatact acctgggttc cgtggctatg caggctgaga agttcccctc
     3481 cgagcccaac cgcctgctgc tgctgcacgg tttcctcgac gagaacgtgc acttcgctca
     3541 cacctctatc ctgctgtcct tcctcgtgcg tgctggcaag ccttacgacc tccagatcta
     3601 cccccaggag cgtcactcca tccgcgtgcc cgagtccggc gagcactacg aactccattt
     3661 gctgcactac ctgcaggaga acctgggctc ccgtattgct gctctaaaag tgatataata
     3721 ataacattgg aagtggataa tctagagggc cctattctat agtgtcacct aaatgctaga
     3781 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     3841 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     3901 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     3961 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     4021 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     4081 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4141 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4201 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4261 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4321 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4381 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4441 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     4501 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4561 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4621 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4681 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4741 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     4801 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     4861 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4921 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4981 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     5041 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     5101 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     5161 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     5221 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     5281 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5341 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5401 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     5461 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5521 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5581 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5641 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5701 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5761 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5821 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5881 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5941 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6001 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     6061 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6121 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6181 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6241 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6301 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6361 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6421 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6481 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6541 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6601 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6661 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6721 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6781 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6841 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6901 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6961 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7021 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7081 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7141 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7201 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7261 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7321 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7381 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7441 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7501 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7561 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7621 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7681 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7741 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7801 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7861 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7921 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7981 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8041 ggcgacacgg aaatgttgaa tactcatact cttccttttt cattattatt gaagcattta
     8101 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8161 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
